Effects in broiler hens of genetic lines differing in fertility, biotin supplementation, and age on relative abundance of oviductal transforming growth factor-ß and carbonic anhydrase mRNA transcripts.

There was evaluation of effects of biotin administration on oviductal abundance of transforming growth factor-ß (TGF-ß) and carbonic anhydrase (CA) mRNA transcript in younger and older broiler hens of relatively lesser and greater fertility lines. Additionally, effects of biotin supplementation on attenuation of age-related subfertility were evaluated. Hens from the relatively greater (Line D, n = 60) and lesser (Line B, n = 60) fertility rate line were randomly assigned to three treatment groups. Biotin was not or was administered in drinking water from 30 to 33 (younger age) and 53 to 56 (older age) wk of age to have access to no biotin (T0), or 0.3 (T1), or 0.45 (T2) mg/L of biotin. There was assessment the relative oviductal abundances of TGF-ß and CA mRNA transcript abundances. Supplemental biotin and age had no effect on the relative abundance of oviductal TGF-ß mRNA transcript in hens of Line D. There, however, was a ten-fold greater abundance of TGF-ß in hens of the T0 group of Line B compared with Line D. Relative abundance of TGF-ß mRNA transcript was greater in younger hens of Line B; however, biotin supplementation of older hens of the T2 group of Line B resulted in a similar TGF-ß abundance to that of younger hens. Inconstant with the TGF-ß abundance, CA abundance in hens of Line B was not affected by supplemental biotin or bird age. Overall, differences in TGF-ß or CA abundances did not affect fertility of broiler hens.

Diet-Induced Obesity Elicits Macrophage Infiltration and Reduction in Spine Density in the Hypothalami of Male but Not Female Mice.

Increasing prevalence in obesity has become a significant public concern. C57BL/6J mice are prone to diet-induced obesity (DIO) when fed high-fat diet (HFD), and develop chronic inflammation and metabolic syndrome, making them a good model to analyze mechanisms whereby obesity elicits pathologies. DIO mice demonstrated profound sex differences in response to HFD with respect to inflammation and hypothalamic function. First, we determined that males are prone to DIO, while females are resistant. Ovariectomized females, on the other hand, are susceptible to DIO, implying protection by ovarian hormones. Males, but not females, exhibit changes in hypothalamic neuropeptide expression. Surprisingly, ovariectomized females remain resistant to neuroendocrine changes, showing that ovarian hormones are not necessary for protection. Second, obese mice exhibit sex differences in DIO-induced inflammation. Microglial activation and peripheral macrophage infiltration is seen in the hypothalami of males, while females are protected from the increase in inflammatory cytokines and do not exhibit microglia morphology changes nor monocyte-derived macrophage infiltration, regardless of the presence of ovarian hormones. Strikingly, the anti-inflammatory cytokine IL-10 is increased in the hypothalami of females but not males. Third, this study posits a potential mechanism of obesity-induced impairment of hypothalamic function whereby obese males exhibit reduced levels of synaptic proteins in the hypothalamus and fewer spines in GnRH neurons, located in the areas exhibiting macrophage infiltration. Our studies suggest that inflammation-induced synaptic remodeling is potentially responsible for hypothalamic impairment that may contribute to diminished levels of gonadotropin hormones, testosterone, and sperm numbers, which we observe and corresponds to the observations in obese humans. Taken together, our data implicate neuro-immune mechanisms underlying sex-specific differences in obesity-induced impairment of the hypothalamic function with potential consequences for reproduction and fertility.

Immunocontraceptive target repertoire defined by systematic identification of sperm membrane alloantigens in a single species.

Sperm competence in animal fertilization requires the collective activities of numerous sperm-specific proteins that are typically alloimmunogenic in females. Consequently, sperm membrane alloantigens are potential targets for contraceptives that act by blocking the proteins' functions in gamete interactions. Here we used a targeted proteomics approach to identify the major alloantigens in swine sperm membranes and lipid rafts, and thereby systematically defined the repertoire of these sperm-specific proteins in a single species. Gilts with high alloantibody reactivity to proteins in sperm membranes or lipid rafts produced fewer offspring (73% decrease) than adjuvant-only or nonimmune control animals. Alloantisera recognized more than 20 potentially unique sperm membrane proteins and five sperm lipid raft proteins resolved on two-dimensional immunoblots with or without prior enrichment by anion exchange chromatography. Dominant sperm membrane alloantigens identified by mass spectrometry included the ADAMs fertilin α, fertilin ß, and cyritestin. Less abundant alloantigens included ATP synthase F1 ß subunit, myo-inositol monophosphatase-1, and zymogen granule membrane glycoprotein-2. Immunodominant sperm lipid raft alloantigens included SAMP14, lymphocyte antigen 6K, and the epididymal sperm protein E12. Of the fifteen unique membrane alloantigens identified, eleven were known sperm-specific proteins with uncertain functions in fertilization, and four were not previously suspected to exist as sperm-specific isoforms. De novo sequences of tryptic peptides from sperm membrane alloantigen "M6" displayed no evident homology to known proteins, so is a newly discovered sperm-specific gene product in swine. We conclude that alloimmunizing gilts with sperm membranes or lipid rafts evokes formation of antibodies to a relatively small number of dominant alloantigens that include known and novel sperm-specific proteins with possible functions in fertilization and potential utility as targets for immunocontraception.

Effect of lycopene on semen quality, fertility and native immunity of broiler breeder.

1. The effect of drinking water supplementation with lycopene on the semen quality, fertility and immunity of broiler breeders was evaluated. 2. Broiler breeder males were individually caged from 25 to 42 weeks old and divided into two group: L group, treated birds (lycopene 0.5 g/l) and C group, control birds. Laying hens were divided into two groups and artificially inseminated. 3. Semen variables were evaluated and daily fertility recorded. Serum bactericidal activity was tested. 4. Semen production and viability were affected by lycopene supplementation. Serum bactericidal activity was better in L than in C group. The fertility rate curve of the L group displayed a positive trend.

A novel testis-specific Na+/H+ exchanger is involved in sperm motility and fertility.

Sodium-hydrogen exchanger as a channel for regulation of intracellular pH might be a crucial modulator of sperm capacitation and motility. Three members of this family have been identified in spermatozoa. A novel protein testis-specific sodium-hydrogen exchanger named mtsNHE was cloned in the present study. The mtsNHE localizing on principle piece of sperm flagellum contained 12 predicted transmembrane regions without cytoplasmic fragment at carboxyl terminus. Hydrophilic region was common in the sodium-hydrogen exchanger family members. Polyclonal antibodies to trans-membrane region significantly reduced sperm motility, acrosome reaction and ratio of in vitro fertilization. By in-pouring the antibodies in sperm solution, intracellular pH and calcium concentration were decreased. Muscle injection of female mice with the specific gene vaccine of mtsNHE, significantly stepped down fertility rate. Considering its specific expression and involvement in the regulation of fertility, the mtsNHE might be a potential target molecule for developing a new male contraceptive.

The differential effect of injecting estradiol-17beta, testosterone, and hydrocortisone during the immune adaptive period on the fertility of female mice.

PROBLEM: Female mice injected with estradiol-17beta (E2) and testosterone during the immune adaptive period are infertile as adults. Study 1 examined the effect of the day of injection of E2 and testosterone on the incidence of infertility in two strains of mice. Study 2 examined the effect of hydrocortisone on E2-induced infertility. METHOD OF STUDY: Study 1: Neonatal (C57BL/6J x A/J)F1 B6A and (C3H/HeJ x 129J)F1 C31 female mice were injected from 0 to 3 and from 3 to 6 days of age with either 20 microg E2 or 20 microg testosterone. Animals were tested for fertility by mating with fertile males. Study 2: Neonatal B6A females were injected with 20 microg E2 with/without 1000 microg hydrocortisone on days 1, 3, 5, 7, and 10. At adulthood, ovaries were examined for the presence of corpora lutea (CLs). RESULTS: Study 1: The incidence of E2-induced infertility in adult B6A and C31 females decreased over three consecutive matings. In contrast, the incidence of testosterone-induced infertility in adult B6A and C31 females increased. E2 caused the highest incidence of infertility in C31 females when injected prior to 3 days of age. In B6A mice, E2 caused the highest incidence of infertility when injected after 3 days of age. Study 2: When hydrocortisone was injected with E2, 90% of the B6A females had ovaries with CLs at 100 days of age. Without hydrocortisone, only 16% of the B6A females injected with E2 had ovaries with CLs. CONCLUSION: Study 1: The incidence of infertility caused by injections of E2 is dependent on the strain of mice and the day(s) injected. The incidence of infertility caused by injections of testosterone is independent of the strain of mice. Study 2: Hydrocortisone prevents E2-induced infertility. It is proposed that injections of E2 during the immune adaptive period alter T-cell maturation, which contributes to E2-induced infertility.